Effect of Lactic acid and P levels on rumen microbial dynamics · 2017. 10. 17. · University of Veterinary Medicine, Vienna (Vetmeduni Vienna) Effect of Lactic acid and P levels

University of Veterinary Medicine, Vienna (Vetmeduni Vienna)

Effect of Lactic acid and P levels on rumen microbial dynamics R.M. Petri, B.U. Metzler-Zebeli & Q. Zebeli

Phosphorus Cycle in Ruminants

Dietary P Fecal P Salivary P

Microbial P

Milk P

(Humer and Zebeli, 2015)

Improving P Availability

Faecal excretion

of P

Efficiency of

phytate P digestion

Brask/Pedersen et al. 2013; Jarret et al. 2014; Humer et al. 2015a

0

20

40

60

80

100

0 2 4 6 8 10 12 14 16 18 20 22 24

In s

itu d

egra

datio

n of

P

(%)

Incubation time (h)

CONLA

*P < 0.05

Khol-Parisini et al. (2015)

Degradation of Organic P +LA

Hypothesis

The treatment of concentrates with 5% lactic acid in early lactation diets will increase the availability

of P to microbal degradation and decrease the faecal excretion of P

Feeding Experiment

Lactating dairy cows (n=17)

4 primiparous; 13 multiparous

Mixed breeds (Average BW 756 ± 159 kg)

Diets Ingredient, % of DM Nutrient composition % of DM

Harder et al. (2015)

Dietary Treatment Item CON LA+P LA-P DM, % of fresh feed 43.1 43.1 42.7 Ash 7.11 7 6.97 CP 18.5 18.9 19.5 NDF 34.8 33.5 31.6 ADF 22.3 21.7 19.7 Ether extract 2.1 2.16 2.22 NFC4 37.5 38.4 39.8 Ca, g/kg DM 9.63 9.66 9.58 P, g/kg DM 4.53 4.64 4.13 NEL, MJ/kg DM 7.06 7.23 7.17

Dietary Treatment Item CON LA+P LA-P Meadow hay 10 10 10 Grass silage 10 10 10 Corn silage 33 33 33 Concentrate with inorganic P, without LA

47 - - Concentrate with inorganic P, with LA*

- 47 - Concentrate without inorganic P, with LA*

- - 47

*soaked with 5% lactic acíd for 24h before feeding

Sample Analysis 50 g faeces per cow were collected rectally

on d 29 and stored at -20°C Chemical Analysis, DNA extraction,

qPCR, Sequencing Illumina MiSeq platform and stitched by

Microsynth (Balgach, Switzerland). QIIME (Caporaso et al. 2010) PiCRUST analysis with KEGG (Langille et

al. 2013) Proc Mixed Procedure of SAS (V.9.4)

Alpha Diversity - Results

Treatment P-value of Contrasts

Variable CON LA P+

LA P- SEM

CON vs. LA

P+ vs. P-

Chao1 6990 6598 6902 118.7 0.521 0.772

Simpson 0.996 0.994 0.995 0.0007 0.317 0.912

Shannon 9.635 9.440 9.518 0.0564 0.346 0.905

2,737,093 quality filtered sequences

9398 Total OTUs 152 OTUs > 0.001% of

sequences 161,005 mean

sequences per sample

OTU Significant Variation: Relative Abundance

OTU‘s Treatment P-value of Contrasts

CON P+LA P-LA SEM CON vs. LA P+ vs. P- OTU 4 0.0113 0.0088 0.0082 0.0010 0.04 0.16 OTU 6 0.0107 0.0080 0.0075 0.0010 0.05 0.20 OTU 47 0.0025 0.0054 0.0035 0.0009 0.005 0.45 OTU 54 0.0017 0.0054 0.0025 0.0011 0.04 0.31 OTU 66 0.0015 0.0028 0.0030 0.0005 0.01 0.10 OTU 73 0.0030 0.0012 0.0022 0.0005 0.04 0.91 OTU 88 0.0027 0.0016 0.0023 0.0003 0.05 0.55 OTU 105 0.0017 0.0024 0.0013 0.0003 0.55 0.05 OTU 123 0.0012 0.0016 0.0025 0.0004 0.05 0.02 OTU 127 0.0028 0.0016 0.0017 0.0004 0.01 0.007 OTU 138 0.0020 0.0010 0.0010 0.0003 0.04 0.26

OTU Classification

OTUs

CON vs. LA

(P-value) P+ vs. P- (P-value) Description NCBI

Percent Similarity

Clostridium Cluster based

on NCBI Description via Green

Genes

OTU 4 0.04 0.16 Fastidiosipila sanguinis strain CCUG 47711 90% iii Ruminococcaceae fam.

OTU 6 0.05 0.20 Thermotalea metallivorans strain B2-1 86% xi Clostridiaceae family

OTU 47 0.005 0.45 Alkalibaculum bacchi strain CP11 89% xv Clostridiales order

OTU 54 0.04 0.31 Saccharofermentans acetigenes strain P6 90% iii Ruminococcaceae fam.

OTU 66 0.01 0.10 Saccharofermentans acetigenes strain P6 91% iii Ruminococcaceae fam.

OTU 73 0.04 0.91 Gracilibacter thermotolerans strain JW/YJL-S1 87% i/ii/iii Clostridiales order

OTU 88 0.05 0.55 Sporobacter termitidis strain SYR 94% iv Ruminococcaceae fam.

OTU 123 0.05 0.02 Defluviitalea saccharophila strain LIND6LT2 88% iii Ruminococcaceae fam.

OTU 127 0.01 0.007 Peptococcus niger strain JCM 6506 89% n/a RC4-4 genus

OTU 138 0.04 0.26 Gracilibacter thermotolerans strain JW/YJL-S1 87% i/ii/iii Clostridiales order

qPCR Treatment P-value Variable CON P+LA P-LA SEM Treat Phos Total Bacteria (log10 Gene Copies/g faeces) 10.90 10.84 10.78 0.034 0.565 0.528

Percent of Total Bacteria determined as bacterial groups or speices %) Genus Prevotella 2.97 3.28 3.04 0.094 0.394 0.680 Lactobacillus group 0.0001 0.0001 0.0002 0.000 0.591 0.210 Clostridium cluster I 0.004 0.004 0.009 0.002 0.570 0.203 Clostridium cluster IV 0.03 0.03 0.02 0.000 0.664 0.620 Clostridium cluster XIVa 3.96 3.82 3.68 0.082 0.602 0.602 Genus Ruminicoccus 6.52 6.03 6.02 0.164 0.249 0.541 Turicibacter spp. 0.0026 0.0023 0.0021 0.000 0.572 0.653 Streptococcus spp. 0.0012 0.0011 0.0008 0.000 0.107 0.080 Total Methanogen (log10 Gene Copies/mL) 0.05 0.05 0.04 0.005 0.102 0.079

KEGG Predictive Pathways Phosphorus Responding OTUs

Contrasts

Control LA +P LA -P SEM CON vs.

LA P+ vs.

P- Environmental Information Processing

Membrane Transport Phosphotransferase system (PTS) 537 319 372 74.8 0.24 0.73

Signal Transduction Phosphatidylinositol signaling system 316 158 190 47.3 0.17 0.64

Metabolism Carbohydrate Metabolism Inositol phosphate metabolism 316 158 190 47.3 0.17 0.64 Carbohydrate Metabolism Pentose phosphate pathway 2771 1400 1675 412 0.17 0.64 Energy Metabolism Oxidative phosphorylation 3664 1740 2101 594 0.18 0.64 Lipid Metabolism Glycerophospholipid metabolism 2251 1163 1386 325 0.17 0.64 Metabolism of Amino Acids

Phosphonate and phosphinate metabolism 69 26 33 15 0.24 0.66

Faecal Phosphorus

Treatment P-Value

Contrasts

(g/kg)

CON LA P+

LA P-

SEM CON vs. LA

P+ vs. P-

Faecal P 10.3 10.5 9.0 0.27 0.36 0.03

Conclusions

LA increases availability of organic P

P was not a limiting factor for faecal microbiota

The use of LA in early lactation diets is a viable option for reducing cost and P pollution

Thanks goes to:

Graduate Students: H. Harder S. Mickdam

Technical Assistance: A. Dockner M. Wild S. Eisen

Scientific Personnel: R. Khiaosa-Ard B. Metzler- Zebeli Q. Zebeli

Funding Agency: